Identification and Functional Analysis of Two Novel Genes-Geranylgeranyl Pyrophosphate Synthase Gene (AlGGPPS) and Isopentenyl Pyrophosphate Isomerase Gene (AlIDI)-from Aurantiochytrium limacinum Significantly Enhance De Novo beta-Carotene Biosynthesis in Escherichia coli
文献类型: 外文期刊
作者: Shi, Shitao 1 ; Chang, Yi 2 ; Yu, Jinhui 3 ; Chen, Hui 2 ; Wang, Qiang 2 ; Bi, Yuping 1 ;
作者机构: 1.Shandong Univ, Sch Life Sci, Qingdao 266237, Peoples R China
2.Henan Univ, Sch Life Sci, State Key Lab Crop Stress Adaptat & Improvement, Kaifeng 475004, Peoples R China
3.Shandong Acad Agr Sci, Inst Crop Germplasm Resources, Jinan 250100, Peoples R China
关键词: Aurantiochytrium limacinum; terpenoid precursor; geranylgeranyl pyrophosphate synthase (GGPPS); isopentenyl pyrophosphate isomerase (IDI); beta-carotene biosynthesis
期刊名称:MARINE DRUGS ( 影响因子:5.4; 五年影响因子:5.5 )
ISSN:
年卷期: 2023 年 21 卷 4 期
页码:
收录情况: SCI
摘要: Precursor regulation has been an effective strategy to improve carotenoid production and the availability of novel precursor synthases facilitates engineering improvements. In this work, the putative geranylgeranyl pyrophosphate synthase encoding gene (AlGGPPS) and isopentenyl pyrophosphate isomerase encoding gene (AlIDI) from Aurantiochytrium limacinum MYA-1381 were isolated. We applied the excavated AlGGPPS and AlIDI to the de novo beta-carotene biosynthetic pathway in Escherichia coli for functional identification and engineering application. Results showed that the two novel genes both functioned in the synthesis of beta-carotene. Furthermore, AlGGPPS and AlIDI performed better than the original or endogenous one, with 39.7% and 80.9% increases in beta-carotene production, respectively. Due to the coordinated expression of the 2 functional genes, beta-carotene content of the modified carotenoid-producing E. coli accumulated a 2.99-fold yield of the initial EBIY strain in 12 h, reaching 10.99 mg/L in flask culture. This study helped to broaden current understanding of the carotenoid biosynthetic pathway in Aurantiochytrium and provided novel functional elements for carotenoid engineering improvements.
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